中国北方汉族人群基因多态性与青少年特发性脊柱侧凸关联研究

发布时间：2018-04-30 17:25

  本文选题：青少年特发性脊柱侧凸 + 遗传学　； 参考：《北京协和医学院》2015年博士论文

【摘要】：研究背景青少年特发性脊柱侧凸(Adolescent Idiopathic Scoliosis, AIS)是所有脊柱侧凸患者中最常见的一类,不同人群发病率为0.47-5.2%。在其病因学研究方面,人们很早就发现特发性测凸患者的一级亲属患病风险明显增高,但经过多年研究,目前尚未明确提出一个致病基因。近年来,国内、国际上有多个研究团队从单核苷酸多态性(Single Nucleotide Polymorphism, SNP)角度着手进行遗传学病因研究,共涉及39个AIS相关候选基因,包括IGF-1、ESR1、LBX-1、 GPR126等。但既往关于AIS的SNP研究存在诸多不足：各个研究中心的样本数量参差不齐,一些小的研究团队仅有数十名患者,导致研究结论可信性低；不同人种之间的结果重复性差,体现了不同种族之间的遗传学背景差异；缺乏大规模中国北方汉族人群研究资料,不能直接应用国外研究的结果；缺乏深入的功能试验,使研究成果停留在“关联性”而不能建立病因学上的因果关系。因此,有必要基于我国人群进行既往研究的验证工作并对重要候选基因进行深入功能试验。研究目的1.建立中国北方汉族人群青少年特发性脊柱侧凸遗传学研究队列；2.在本队列中验证既往报道中与AIS发病相关的SNP位点；3.针对重点候选基因,通过深入功能分析,尝试解释其致病机制。研究方法确立入组及排除标准,收集于北京协和医院骨科行手术治疗的AIS患者外周血样本及临床资料,建立研究队列。通过检索、回顾文献,确定需要验证的AIS相关基因及SNP位点。根据病例-对照研究策略,使用Sequenom MassArray分型平台,对患者及正常对照的候选SNP进行基因分型,并应用PLINK及SPSS软件进行统计分析。针对重点基因LBX1,应用候选基因关联分析法选取基因及其周围的SNP位点,通过Sequenom MassArray技术进行分型,得到功能试验候选位点。针对该位点,使用双荧光报告素酶系统研究其对LBX1表达的影响,最终给出致病机理模型。研究结果1.本研究共纳入中国北方汉族AIS患者180例,性别、民族匹配的正常对照182例；2.通过文件回顾,共纳入rs1065755等20处AIS相关SNP位点；3. rs11190870、rsl2885713和rs2300500三个位点在等位基因关联分析、基因型关联分析以及遗传模型分析中均与AIS发病相关,而rs6570507、 rs4753426仅在遗传模型分析中与AIS具有相关性,分别符合隐形模型及趋势模型；4.对LBX1基因应用候选基因多态性分析方法,得到3个与AIS发病相关SNP位点：rs11190870,rs1322331及rs625039。其中rs1322331的可信性最高、可进行功能试验；5.通过双荧光报告素酶系统检验,在体外条件下,rs1322331位点为GG纯合型时对下游基因的表达起显著增强作用,而TT纯合型则对下游基因的表达无显著作用。结论1.中国北方汉族人群中,既往文献报道的rs11190870、rsl2885713.rs2300500、 rs6570507和rs4753426等位点与AIS发病具有相关性；2.中国北方汉族人群中,LBXl基因处与AIS发病相关联且可信度最高的SNP位点是rs1322331；3.GG基因型的rs1322331可上调其所在的启动子区域功能,使下游基因表达上调。据此,我们提出LBX1过量表达导致AIS发病的假说。
[Abstract]:Background adolescent idiopathic scoliosis (Adolescent Idiopathic Scoliosis, AIS) is the most common category in all scoliosis patients. The incidence of different populations is 0.47-5.2%. in its etiology research. People have found a significant increase in the risk of first-degree disease in patients with idiopathic protruding, but after years of study, the number of patients with idiopathic scoliosis was found to be significantly higher. In recent years, a number of research teams in the world have conducted genetic studies on Single Nucleotide Polymorphism (SNP), involving 39 AIS related candidate genes, including IGF-1, ESR1, LBX-1, GPR126 and so on. However, there are many previous SNP studies on AIS. There are many shortcomings: the sample size of each research center is uneven, and some small research teams only have dozens of patients, which leads to low credibility of the research conclusions, poor reproducibility of the results between different races, reflecting the genetic background difference between different races, and the lack of a large scale in the northern Han population. With the results of foreign studies, the lack of in-depth functional tests makes the research results stay in "relevance" and can not establish causality in the etiology. Therefore, it is necessary to verify the previous research and test the important candidate genes based on the population of our country. Objective 1. to establish the Han population in Northern China. A cohort of adolescent idiopathic scoliosis genetic studies; 2. in this cohort, the SNP loci associated with AIS were verified in this cohort; 3. for key candidate genes, the pathogenesis was explained by in-depth functional analysis. The research methods established the group and exclusion criteria and collected in the surgical treatment of the Department of orthopedics in Peking Union Medical College Hospital, AIS The patient's peripheral blood samples and clinical data were set up. The AIS related genes and SNP loci were identified by retrieving and reviewing the literature. According to the case-control study strategy, the Sequenom MassArray typing platform was used to classify the patients and the normal control candidate SNP, and the PLINK and SPSS software were used for statistical analysis. In view of the key gene LBX1, the candidate gene association analysis method was used to select the gene and its surrounding SNP loci, and the functional test candidate loci were obtained by Sequenom MassArray technique. In view of the loci, the effect of the double fluorescent reporter system on the expression of LBX1 was studied, and the pathogenic mechanism model was finally given. 1. This study included 180 AIS patients in the Han nationality in northern China, 182 cases of normal control of gender and national match; 2. through document review, a total of 20 AIS related SNP loci were included and three loci of 3. rs11190870, rsl2885713 and rs2300500 were associated with allele correlation analysis, genotype association analysis and genetic model analysis with AIS hair. The disease was related, and rs6570507, rs4753426 only correlated with AIS in the genetic model analysis, which conformed to the stealth model and the trend model respectively. 4. the candidate gene polymorphism analysis method of the LBX1 gene was used to obtain the 3 SNP loci associated with the pathogenesis of AIS: rs11190870, rs1322331 and rs625039., the credibility of rs1322331 was the highest, and the work could be done. Can test; 5. through the double fluorescent reporter system test, under the condition of in vitro, the rs1322331 site is GG homozygous, the downstream gene expression is significantly enhanced, while the TT homozygous type has no significant effect on the downstream gene expression. Conclusion 1. in the Han population of northern China, rs11190870, rsl2885713.rs2300500, R reported in the literature. The loci of s6570507 and rs4753426 are related to the pathogenesis of AIS; 2. in the Han population of northern China, the SNP locus associated with the pathogenesis of AIS and the highest reliability of AIS is rs1322331; the rs1322331 of the 3.GG genotype can increase the function of its promoter region and up-regulation the expression of the downstream genes. Accordingly, we propose that LBX1 overexpression. The hypothesis that causes the onset of AIS.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R682.3
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本文编号：1825488